Overview

This dissertation, Development of Antibody and Antigen Detection Assays and Vaccines for SARS Associated Coronavirus by Hiu-ling, Beatrice, Wong, 黃曉靈, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled Development of antibody and antigen detection assays and vaccines for SARS Associated Coronavirus Submitted by Wong Hiu Ling Beatrice for the degree of Doctor of Philosophy at The University of Hong Kong in December 2007 In the first part of my study, recombinant enzyme-linked immunosorbant assay (ELISA) for antibody and antigen measurement was developed for laboratory diagnosis of severe acute respiratory syndrome (SARS) and seroepidemiology studies. For detection of antibodies, both recombinant SARS coronavirus (SARS-CoV) nucleocapsid (N) protein and spike (S) polypeptide based ELISA were developed. The sensitivities of N-based ELISA are 94.3%, 59.4% and 60.4% respectively for detection of immunoglobulin (Ig) G, IgM and IgA, with respective specificities of 95.3%, 96.6% and 96.6%; while the specificities of S-based ELISA for IgG and IgM detection were 98.6% and 93.9% respectively, with 2 corresponding sensitivities of 58.9% and 74.7%. Seroepidemiology studies were performed using the N-based ELISA with confirmation by S polypeptide Western blot assay. Three of the 400 healthy blood donors who donated blood during the SARS outbreak and 1 of 131 non-pneumonic pediatric inpatients were positive for IgG antibodies. As for antigen detection, capturing ELISAs for detection of N protein in SARS patients were developed, with specificities of 96.7%, 99%, 96% and sensitivities of 52%, 5%, 55% for nasopharyngeal aspirate, urine and fecal specimens respectively. Comparison between quantitative real-time polymerase chain reaction (qRT-PCR) and polyclonal and monoclonal antibody-based N antigen capturing ELISAs showed that the sensitivity of qRT-PCR is higher than those of both N antigen capturing ELISAs. In the second part of my study, the longitudinal IgG, IgM and IgA antibody profiles in patients with pneumonia due to SARS-CoV were studied using serum samples serially collected up to day 240 after the onset of illness. The median times of seroconversion for IgG, IgM and IgA detection were 17, 20.5 and 17 days after disease onset respectively. One, 4 and 1 of the 6 patients who died did not produce any IgG, IgM and IgA antibodies against the nucleocapsid protein of SARS-CoV respectively. 3 In the third part of my study, the cross-reactivity of the N-based ELISA between SARS-CoV and serum samples positive for antibody against human coronavirus-229E (HCoV-229E) or HCoV-OC43 was studied. Three of the 21 and 1 of the 7 convalescent-phase serum samples from persons with antibodies against HCoV-OC43 and HCoV-229E respectively were tested positive by the recombinant SARS-CoV N protein-based ELISA. None of these samples were found to contain a specific antibody in the recombinant SARS-CoV S polypeptide-based Western blot assay. In the fourth part of my study, several different potential vaccines were developed to investigate their ability to generate neutralizing antibodies against SARS-CoV. Sera of mice immunized with intramuscular (i.m.) tPA-optimize800 DNA vaccine (tPA-S-DNA), i.m. CTLA4HingeSARS800 DNA vaccine (CTLA4-S-DNA), polypeptide, oral live-attenuated Salmonella typhimurium that contained CTLA4HingeSARS800 DNA vaccine (Salmonella-CTLA4-S-DNA) and oral Salmonella-CTLA4-S-DNA boosted with intraperitoneal (i.p.) S-polypeptide showed neutralizing antibody titers of

Full Product Details

9781374664449

Table of Contents

Reviews

Author Information

Tab Content 6

Customer Reviews

Recent Reviews

Countries Available